■周期性血小板减少症(CTP)是一种罕见的血液疾病,其特征是血小板计数的周期性波动。CTP通常出现在绝经前妇女身上,这些血小板的波动与月经周期同步。CTP是一种异质性疾病,致病机制尚不清楚。因此,它对探索血小板计数波动与激素周期的关系具有重要意义。
■首先,根据赫尔辛基宣言,我们清洗了健康志愿者的血小板。流式细胞仪用于测量线粒体内部跨膜电位(ΔkWm)去极化,PS暴露,P-选择素表达,和血小板中的GPIIb/IIIa激活。此外,westernblot检测相关蛋白的表达。相应的测定试剂盒测量胱天蛋白酶-3和PDE3A活性。最后,流式细胞术测定用钙黄绿素标记的小鼠血小板。
■我们发现CTP患者的血小板计数与血清雌二醇(E2)水平之间存在反向关系。我们证明E2在体外诱导血小板凋亡,在体内诱导血小板清除。我们进一步发现E2激活磷酸二酯酶3A,抑制蛋白激酶A(PKA),导致PKA介导的血小板凋亡。PKA的激活可保护血小板免受E2诱导的血小板减少症的影响,并增加了小鼠循环血小板的数量。
■我们发现E2通过PDE3A介导的PKA抑制诱导血小板凋亡和清除。PKA的激活挽救了E2诱导的小鼠血小板减少症。因此,我们的研究揭示了E2相关CTP的发病机制,并提出了有希望的治疗策略.
UNASSIGNED: Cyclic thrombocytopenia (CTP) is a rare blood disorder characterized by periodic fluctuations in platelet counts. CTP usually appears in pre-menopausal women, and these fluctuations of platelets are in phase with the menstrual cycle. CTP is a heterogeneous disease, and the pathogenic mechanism is still unclear. Therefore, it harbors great significance for exploring the association of fluctuations in platelet counts with hormonal-cycle.
UNASSIGNED: Firstly, we washed human platelets from healthy volunteers following the Declaration of Helsinki. Flow cytometer was employed to measure the mitochondrial inner transmembrane potential (ΔΨm) depolarization, PS exposure, P-selectin expression, and GPIIb/IIIa activation in platelets. In addition, western blot detected the related protein expression. The corresponding assay kit measured the caspase-3 and PDE3A activity. Finally, flow cytometry determined mouse platelets labeled with calcein.
UNASSIGNED: We find a reverse relationship between the platelet count and serum estradiol (E2) level in a CTP patient. We demonstrated that E2 induces platelet apoptosis in vitro and platelet clearance in vivo. We further discovered that E2 activates phosphodiesterase 3A, which inhibits protein kinase A (PKA), leading to PKA-mediated platelet apoptosis. Activation of PKA protected platelets from E2-induced thrombocytopenia and elevated the number of mice circulatory platelets.
UNASSIGNED: We find that E2 induces platelet apoptosis and clearance through PDE3A-mediated PKA inhibition. Activation of PKA rescues E2-induced thrombocytopenia in mice. Thus, our study reveals a pathogenesis of E2-related CTP and suggests promising therapeutic strategies for the disease.